Eliminating Matrix Effects in Mass Spectrometry Applications

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Erin Chambers, Ziling Lu, Diane M. Diehl and Jeffrey R. Mazzeo [Waters]
EAS 2005
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Rat Plasma
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Interference of the mass spectrometer response from residual matrix components is the definition of matrix effects. Previously, we have shown how different sample matrices, different measuring techniques, and different sample preparation techniques such as SPE and protein precipitation contribute to matrix effects. We continued this research by examining fast gradients vs. slow gradients, a wider range of analytes including polar analytes (most likely to co-elute with matrix salts and polar interferences), and non-polar analytes (most likely to co-elute with matrix hydrophobic interferences). We looked at the amount of matrix remaining in the extracts by running the samples using LC/MS/MS (electrospray ionization) under both scan mode and multiple reaction monitoring. For hydrophobic analytes chromatographed under fast gradients, matrix effects are the most detrimental. Under these conditions, sample preparation is crucial. Protein precipitation is the least effective sample clean-up procedure, resulting in nearly an 80% loss of signal, while a mixed-mode SPE procedure results in no loss of signal. Additionally, we have investigated the degree of matrix effect using various sample preparation techniques and UPLC™ with mass spectrometry. While the level of signal suppression is less using UPLC, effective sample preparation is still necessary to ensure the most robust methods.

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